交联壳聚糖微球偶联胰蛋白酶研究  

Studies on Cross Linked ChitosanMicro-Sphere Immobilizing Trypsin

作　　者：沈明才[1] 谭竹钧[1] 李宇光[1] 

机构地区：[1]广东工业大学轻工化工学院,广东广州510090

出　　处：《药物生物技术》2008年第1期20-23,共4页Pharmaceutical Biotechnology

基　　金：广东工业大学人才基金(编号:052056);广东省自然科学基金项目(No.04010984)

摘　　要：采用反相悬浮交联法,制备交联壳聚糖微球(CCTS),再以戊二醛为偶联剂,偶联胰蛋白酶.研究了胰蛋白酶偶联的最佳条件和偶联后酶的理化性质。结果表明:用终浓度为0.6%的戊二醛,在50℃下,先处理CCTS 6 h,再置冰浴加酶为12 mg/g,搅拌24 h,固定化酶活性回收率达63.4%。胰蛋白酶与壳聚糖微球偶联的最适温度为80℃、最适pH值为7、表观米氏常数(Km)为3.16 mmol/L。与壳聚糖载体相比,CCTS具有机械强度好、粒度均匀、耐酸性能好、对牛血清白蛋白非特异性吸附较弱的优点。A cross-linking chitosan micro-sphere（CCTS） was prepared through inverse suspension cross-linking, and then it was used to immobilize trypsin with glutaraldehyde. The optimum conditions of immobilizing trypsin and the characteristics of immobilizing trypsin were studied in this article. The results indicated that （1）The CCTS reacted with glutaraldehyde whose final concentration was 0. 6% at 50℃ for 6 hours, after adding trypsin 12 mg/g and reacted for 24 hours at 0--5℃. （2）The optimum temperature of immobilized trypsin was 80℃ ,and the pH was about 7. The apparent Michcalis Constant of immobilized trypsin （Km） was 3. 16 mmol/L. Compared with the chitosan, the CCTS had the advantage of good strength, symmetrical granularity, and strong acid-resistance. The CCTS had no special adsorption to BSA at the same time.

关 键 词：交联壳聚糖 胰蛋白酶 固定化酶 

分 类 号：R943[医药卫生—药剂学]